KR101973907B1 - Phamaceutical composition for preventing or treating of specific cancers containing extracts, fractions or sargachromenol from Sargassum serratifolium - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for prevention or treatment of certain cancers containing extracts, fractions or sagarchromenol of Tootsumabum extract according to the present invention, wherein the ethanol extract, hexane fraction or sagar chromenol according to the present invention has cytotoxicity The cytotoxicity to certain cancer cells (particularly glioblastoma and osteosarcoma cancer) is very high, and thus may be useful as a therapeutic agent for a specific cancer.

Description

FIELD OF THE INVENTION The present invention relates to a pharmaceutical composition for prevention or treatment of certain cancers, especially fractions or sargachromenol from sargassum serratifolium,

The present invention relates to a pharmaceutical composition for preventing or treating certain cancers containing extracts, fractions or sagarchromenols.

Glioblastoma, one of the malignant brain gliomas, refers to the fact that the cells of the brain are transformed into malignant tumors. Although the resection is the most commonly used treatment for malignant brain gliomas, it is not possible to distinguish it from the surrounding normal tissue due to the specificity of the lesion site and the strong invasion of the brain glioma. Therefore, besides the treatment by resection, This is an additional situation.

Osteosarcoma is a primary malignant tumor originating from the bone and is the most common disease in the teenage period. Currently, osteosarcoma is a treatment for surgery and antimicrobial therapy, but after 5 years the survival rate is as low as 20-30%, the need for new therapeutic drugs and techniques.

Accordingly, the present inventors have found that the ethanol extract of Sargassume serratifolium , the fraction of hexane and the Sargachromenol isolated therefrom are highly cytotoxic to certain cancer cells while having no cytotoxicity to normal cells, .

Published Patent Application No. 10-2004-0092302

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer comprising an extract of Sargassume serratifolium as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises a fraction obtained from an ethanol extract of Sargassume serratifolium as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises, as an active ingredient, sagachromenol represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for preventing or treating cancer, which comprises an extract of Sargassume serratifolium as an active ingredient. Herein, the extract may be an ethanol extract, and the cancer is gastric cancer, glioblastoma, cervical cancer, or liver cancer, preferably gastric cancer, glioblastoma or liver cancer, more preferably glioblastoma.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, which comprises a fraction obtained from an ethanol extract of Sargassume serratifolium as an active ingredient. Here, the fraction is an ethyl acetate fraction or a hexane fraction, and preferably a hexane fraction. The cancer is gastric cancer, glioblastoma, cervical cancer, or liver cancer, preferably gastric cancer, glioblastoma, or liver cancer, more preferably glioblastoma.

Further, the present invention provides a pharmaceutical composition for preventing or treating cancer, which comprises sargachromenol represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 1]

Herein, the cancer is gastric cancer, glioblastoma, liver cancer, or osteosarcoma, preferably glioblastoma, liver cancer, or osteosarcoma, more preferably osteosarcoma.

The ethanol extract, hexane fraction or sagar chromenol according to the present invention has a very high cytotoxicity against certain cancer cells (especially glioblastoma and osteosarcoma) without any cytotoxicity against normal cells, . ≪ / RTI >

FIG. 1 is a graph showing the survival rate of various cancer cells of Sargachromenol isolated from sawtooth moths in Production Example 2. FIG.
FIG. 2 is a graph showing the survival rate of osteosarcoma cancer cells of Sargachromenol isolated from sawtooth moths in Production Example 2. FIG.
Fig. 3 is a Western blot evaluation image showing the expression level of the cell cycle control-related protein of Sargachromenol isolated from sawtooth moths in Production Example 2. Fig.
4 is a graph showing cytotoxicity of the ethyl acetate fraction of Production Example 1-2 for four cancer cell lines.
5 is a graph showing cytotoxicity of four kinds of cancer cell lines of the ethanol extract of Preparation Example 1-1.
6 is a graph showing cytotoxicity of the hexane fraction of Production Example 1-2 to four cancer cell lines.
FIG. 7 is a graph showing cytotoxicity observed when the ethanol extract of Preparation Example 1-1 and the hexane fraction of Preparation Example 1-2 were treated to human pneumatocyte (U87MG) by concentration. FIG.
FIG. 8 is a graph showing cytotoxicity observed when the hexane fraction of Production Example 1-2 was treated by concentration in human pseudomyocyte (U87MG) and normal kidney (HEK293) cells.
FIG. 9 is an image obtained by Western blot analysis of the expression amount of MAPK (Mitogen Activated Protein Kinase) after treating the fraction of hexane of Production Example 1-2 with human pseudoepithelial cell (U87MG).
FIG. 10 is an image obtained by treating the fraction of hexane of Production Example 1-2 with human pseudomyocyte (U87MG) followed by immunofluorescence staining.
FIG. 11 is an image obtained by observing the cytoskeleton of human pseudomyxoma cell line (U87MG) by an inverted microscope when the hexane fraction of Production Example 1-2 was treated.
FIG. 12 is an image obtained by western blotting the expression amount of a factor necessary for the cytoskeleton formation of human pneumocyte subpopulation (U87MG) when treating the hexane fraction of Production Example 1-2.
FIG. 13 is an image obtained by treating the human pseudoepancreatic cell line (U87MG) with the hexane fraction of Production Example 1-2 and confirming invasiveness through a Transwell invasion assay.
14 is an image obtained by Western blotting the amount of expression of MMP-2 and MMP-9 after treating the hexane fraction of Production Example 1-2 with human pseudomyocyte (U87MG).
FIG. 15 is an image obtained by treating the human pseudomonas cell line (U87MG) with the hexane fraction of Production Example 1-2 and confirming the mobility through the Wound healing assay.
16 is a chromatogram showing four fractions obtained by dissolving n-hexane fraction obtained from ethanol extraction into methanol and using HPLC.
17 is a chromatogram obtained by re-chromatography of the fraction No. 2 in Fig. 16 to obtain three secondary fractions.
18 is a chromatogram obtained by re-chromatography of the fraction No. 2 in Fig. 17 under the same conditions again.

Pharmaceutically acceptable salts

Sargachromenol according to an embodiment of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, acid addition salt formed by a pharmaceutically acceptable free acid is useful. The term pharmaceutically acceptable salt means a concentration that is relatively non-toxic to a patient and has an innocuous effective action, and that side effects caused by the salt are not adversely affected by any of the basic compounds of sagarchromenol which does not degrade the beneficial effects of the base compound of sagarpronone Organic or inorganic addition salts.

Examples of the inorganic acid include hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid and phosphoric acid. Examples of the organic acid include citric acid, acetic acid, lactic acid, maleic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, tartaric acid, succinic acid, malonic acid, succinic acid, malonic acid, glutamic acid, aspartic acid, oxalic acid, P-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid.

These salts also include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, the acid addition salt may be selected from the group consisting of acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camylate, citrate, eddylate, Hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, malate, glucoside, gluconate, gluconate, glucuronate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / Hydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydrogen phosphate, dihydroxyacetate, Lactate, stearate, succinate, tartrate, tosylate, trifluoroacetate Diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, and the like. Preferred is hydrochloride or trifluoroacetate.

The acid addition salt according to the present invention can be obtained by a conventional method, for example, by dissolving sagarpromone in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., Or may be prepared by evaporating the solvent and excess acid under reduced pressure, followed by drying or crystallization in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Further, the present invention encompasses all of the above-mentioned solvates, hydrates, isomers, optical isomers, and the like, which can be prepared therefrom, as well as the above-mentioned sagarginol and its pharmaceutically acceptable salts.

Pharmaceutical composition

The compound of the present invention may be administered orally or parenterally in various dosage forms during clinical administration. In the case of formulation, the diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, .

Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches and the like, which may contain one or more excipients such as starch, calcium carbonate, It is prepared by mixing sucrose, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like are included in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

The effective dose of the compound of the present invention on the human body may vary depending on the age, weight, sex, dosage form, health condition and disease severity of the patient, and is generally about 0.001-100 mg / kg / 0.0 > mg / kg / day. ≪ / RTI > It is generally from 0.07 to 7000 mg / day, preferably from 0.7 to 2500 mg / day, based on an adult patient weighing 70 kg, and may be administered once a day It may be divided into several doses.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

≪ Preparation Example 1 > Sargassume serratifolium ) Preparation of Extracts and Fractions

<1-1> Preparation of ethanol extract

The sawtooth moths used in the present invention were obtained by scuba diving with natural acids spontaneously grown in Gijang-gun, Busan City, and the species identification of Kijangsan was carried out by Prof. Choi Chang-geun of Ecological Engineering Department, Pukyong National University. 2 kg of the powder obtained by naturally drying in a dark place after being dried in a sunny place was placed in 8 L of alcohol (95% ethanol), and the mixture was refluxed and cooled for 3 hours at 70 캜 for 3 hours. Thereafter, the foreign substance was removed with an ultrafilter (Nitodenko, Japan) Respectively. The filtrate extract was 200 ~ 25g of crude extract as an extracted residue after removing the solvent at 40 [deg.] C with a vacuum rotary evaporator. The thus obtained Saw Moth extract was used as a sample in the following experiment while being stored at 4 ° C in a refrigerator.

<1-2> Preparation of n-hexane, ethyl acetate, butanol and water-soluble fractions from ethanol extracts of tegument

50 g of the ethanol extract of Tooth Moth (ethanol) prepared in Preparation Example 1-1 was dissolved in 1 L of a mixed solvent of water and ethanol (9: 1, v / v), the mixture was poured into a separating funnel, , The separating funnel was shaken and then equilibrated. The upper n-hexane-soluble fraction was collected, treated with sodium sulfate (anhydrous), filtered and concentrated. By repeating this process five more times, 42 g of n-hexane fraction was obtained.

After fractionation with n-hexane, the remaining fraction (H ₂ O layer) was equilibrated with ethyl acetate and the separatory funnel was shaken and equilibrated. The upper ethyl acetate soluble fraction was collected and concentrated to obtain an ethyl acetate fraction.

The same amount of butanol was added to the remaining fraction (H ₂ O layer) after the ethyl acetate fraction, and the separatory funnel was shaken and then equilibrated. Then, the butanol-soluble portion of the upper layer was collected and concentrated to obtain a butanol fraction, and the remaining water-soluble fraction was used as a water fraction.

The ethyl acetate fraction (1L), butanol fraction (1L) and water fraction (0.9L) thus obtained were concentrated under reduced pressure at 40 ° C in a vacuum rotary condenser to remove the solvent, and then 1.2 g, 2.1 g, 4.6 g each was finally obtained as ethyl acetate fraction, butanol fraction and water fraction. Each of the fractions thus obtained was used as a sample for the following experiment while being stored in a refrigerator at 4 캜.

&Lt; Preparation Example 2 > Sargassume serratifolium ) To isolate Sargachromenol (SCM)

12.2 g of the n-hexane fraction obtained in Preparation Example 1-2 was dissolved in 120 mL of methanol, filtered through a membrane filter, and 200 μL was injected to obtain a Phenomenex C18 (2) (15 μm, 21.2 mm × 250 mm) And separated into four fractions by the attached HPLC (see FIG. 16).

The fraction 2 of Figure 16 was rechromatographed using the Example HPLC apparatus and column. The chromatographic conditions were A / B (92.2 / 7.8) to A / B (100/0) for the first time in methanol (A) / water (B) (91.5 / After washing for 6 minutes with A / B (100/0) for 2 minutes, the column was equilibrated with A / B (91.5: 8.5) for 10 minutes. The peak was detected at 270 nm and the flow rate was fixed at 7 mL / min. Chromatography was performed 50-100 times by automatic sample injection with Autosampler to obtain a second fraction of a large fraction of fraction No. 2 (see FIG. 17).

As shown in FIG. 17, the fraction No. 2 showed three peaks, and the peak No. 2 (Frc2-2) was purified again by re-chromatography (126 mg) under the same conditions, and the column for analysis (Phenomenex C18 ), 3 占 퐉, 3 mm 占 150 mm), and it was found to be a single substance (see Fig. 18). The final product was analyzed by NMR and found to be sargachromenol (SCM).

<Experimental Example 1> Sargassume serratifolium Cancer cell survival rate of Sargachromenol (SCM)

The following experiment was carried out to determine whether the sagarchromone prepared in Preparation Example 2 showed toxicity to human cancer cells.

Specifically, normal human skin cells (HaCaT), normal kidney cells (HEK293), normal vascular endothelial cells (HUVEC), human gastric cancer cells (MKN-28, AGS), human glioblastoma (U87MG) 1) and human osteosarcoma cancer cells (MG-63, Saos-2) were divided into 5 × 10 ³ to 1 × 10 ⁴ cells / well on a 96-well plate. (WST-1 solution: 10 μL). After 4 hours, the absorbance was measured at 460 nm using a microplate reader. The measurements were repeated three times and the results were made using an average value, which is shown in FIG.

FIG. 1 is a graph showing the survival rate of various cancer cells of Sargachromenol isolated from sawtooth moths in Production Example 2. FIG.

As shown in FIG. 1, sagachromenol isolated from the sawtooth moth exhibited almost no toxicity in normal cells (HaCaT, HEK293, HUVEC), but showed toxicity in cancer cells. In particular, it showed high cytotoxicity in human osteosarcoma cancer cells (MG-63 and Saos-2), and showed the highest cytotoxicity in Saos-2 cells among them. On the other hand, cytotoxicity was low in gastric cancer cells (MKN-28, AGS), and anticancer efficacy was shown only in specific cancer.

Meanwhile, in order to further examine the anticancer activity of sagarchromenol in human osteosarcoma cancer cells (Saos-2) showing excellent anticancer activity from the results shown in Fig. 1, And the results are shown in Fig.

FIG. 2 is a graph showing the survival rate of osteosarcoma cancer cells of Sargachromenol isolated from sawtooth moths in Production Example 2. FIG.

As shown in FIG. 2, sagarchromenol exhibited a 50% cytotoxic concentration of about 6 μM on osteosarcoma cancer cells, showing high anticancer activity against osteosarcoma cancer cells.

<Experimental Example 2> Evaluation of anticancer efficacy of sargachromenol (SCM) on human osteosarcoma cancer cells

In Experimental Example 1, it was confirmed that Sagar Chromenol exhibited the highest cytotoxicity in human osteosarcoma cancer cell Saos-2. Thus, in Experimental Example 2, the expression level of the cell cycle control-related protein was confirmed by Western blot analysis in order to examine the cytotoxicity of sagarchromenol in Saos-2 cells.

Fig. 3 is a Western blot evaluation image showing the expression level of the cell cycle control-related protein of Sargachromenol isolated from sawtooth moths in Production Example 2. Fig.

As shown in FIG. 3, it was confirmed by Western blot analysis that sagarchromenol decreased the expression levels of CDK 2, 4, 6, Cyclin B 1 and P70S6Kinase, which are related to the cell cycle regulation of Saos-2 cells. In addition, the expression levels of P21, P27, and p-Cdc2 (Tyr 15), which are the cell cycle arrest proteins, were increased. The results showed that Sargachromenol isolated from the sawtooth moth cells had an effect of suppressing the cell cycle by controlling the expression of cell cycle regulating proteins in human osteosarcoma cancer cell Saos-2.

&Lt; Experimental Example 3 > Sargassume serratifolium ) Cancer of extracts and fractions Cytotoxicity Assessment

In Experimental Examples 1 and 2, cytotoxicity of Saga chronomen isolated from the sawtooth moth cells to cancer cells was confirmed. Thus, in Experimental Example 3, it was confirmed that the ethanol extract of Tooth mackerel (Production Example 1-1), the ethyl acetate fraction (Production Example 1-2) and the hexane fraction (Production Example 1-2, HES) The following experiment was conducted to find out.

Specifically, the cells were treated at 50, 100, and 200 μg / mL concentrations in human gastric cancer cells (MKN-28), human cervical cancer cells (HeLa), human glioblastoma (U87MG) The survival rate of the cells was confirmed using WST-1 solution. Each cell was seeded in a 96-well plate at 1 × 10 ⁴ cells / well. The extracts and fractions prepared in Preparation 1-3 were treated for 24 hours at the indicated concentrations. Subsequently, the medium was replaced with medium containing WST-1 solution (100 μl of culture medium, 10 μl of WST-1 solution), and the absorbance was measured at 460 nm using a microplate reader 4 hours later. The measurements were repeated three times and the average values were used to obtain the results.

4 is a graph showing cytotoxicity of the ethyl acetate fraction of Production Example 1-2 for four cancer cell lines.

5 is a graph showing cytotoxicity of four kinds of cancer cell lines of the ethanol extract of Preparation Example 1-1.

6 is a graph showing cytotoxicity of the hexane fraction of Production Example 1-2 to four cancer cell lines.

FIG. 7 is a graph showing cytotoxicity observed when the ethanol extract of Preparation Example 1-1 and the hexane fraction of Preparation Example 1-2 were treated to human pneumatocyte (U87MG) by concentration. FIG.

FIG. 8 is a graph showing cytotoxicity observed when the hexane fraction of Production Example 1-2 was treated by concentration in human pseudomyocyte (U87MG) and normal kidney (HEK293) cells.

As shown in Figs. 4-8, it was confirmed that the hexane fraction of Preparation Example 3 showed relatively high cancer cytotoxicity compared to the ethanol extract of Preparation Example 1 and the ethyl acetate fraction of Preparation Example 2. In addition, the hexane fraction of Preparation Example 3 showed the highest toxicity in human glioblastoma (U87MG) among the four kinds of human cancer cells. In addition, in FIG. 8, it was found that the normal kidney cells (HEK293) show little toxicity and only the human pseudomyxoma cell line (U87MG) shows toxicity. This suggests that the hexane fraction of teaspoon may be useful as a therapeutic agent for human bronchial osteoblast (U87MG).

<Experimental Example 4> Evaluation of cancer cell proliferation inhibition (anti-proliferation) efficacy of hexane fraction of teaspoon moth

Experimental Example 4 was carried out as follows to investigate the cytotoxic effect of the hexane fraction of teat moth, which was confirmed in Experimental Example 3, on human pneumatocytic seedling cells (U87MG) through a change in the intracellular mechanism.

Specifically, in the case of U87MG, the Epidermal Growth Factor Receptor (EGFR), which is one of Receptor Tyrosine Kinases present in the extracellular membrane, is overexpressed by mutation. It is known that overexpression activates Ras / Raf / Mek / Erk (Erk MAPK), one of the mitogen activated protein kinases (MAPKs) in the cell, and indiscriminate proliferation and metastasis of cancer cells. Western blot analysis (FIG. 9) and fluorescent antibody method (Immunofluorescence staining, FIG. 10) were performed to examine the effect of the hexane fraction of the sawtooth moth on Erk MAPK mechanism.

FIG. 9 is an image obtained by Western blot analysis of the expression amount of MAPK (Mitogen Activated Protein Kinase) after treating the fraction of hexane of Production Example 1-2 with human pseudoepithelial cell (U87MG).

FIG. 10 is an image obtained by treating the fraction of hexane of Production Example 1-2 with human pseudomyocyte (U87MG) followed by immunofluorescence staining.

As shown in FIG. 9-10, it was found that the decrease in the expression level of Ras, which is an upper factor of Erk MAPK, and the decrease in the phosphorylation of phosphorylation residues of c-Raf / Mek / Erk, respectively. This confirms that the activity of the Erk MAPK mechanism is reduced. Immunofluorescence staining confirmed that the activity of Erk, a major protein of Erk MAPK, was decreased.

<Experimental Example 5> Inhibitory effect of hexane fraction of teat moth on cytoskeleton formation

Cancer cells continue to form and maintain their own cellular cytoskeletons for indiscriminate proliferation and metastasis. Therefore, if cancer cells can destroy the cytoskeleton or inhibit the process of new formation, it becomes a new strategy to induce the death of cancer cells. Western blot analysis and cell morphology using an inverted microscope were carried out to investigate the effect of Sawtooth mildew extract on the cytoskeleton formation of human glioblastoma (U87MG).

FIG. 11 is an image obtained by observing the cytoskeleton of human pseudomyxoma cell line (U87MG) by an inverted microscope when the hexane fraction of Production Example 1-2 was treated.

FIG. 12 is an image obtained by western blotting the expression amount of a factor necessary for the cytoskeleton formation of human pneumocyte subpopulation (U87MG) when treating the hexane fraction of Production Example 1-2.

As shown in FIG. 11-12, it was confirmed that as the treatment concentration of the hexane fraction of the sawtooth moth increased, the outer shape of the cells became circular compared to the control group. In addition, it was confirmed that the expression of Integrin β1, PI3K p110α, Rac1 / 2/3, WAVE-2, and Arp-2, which are necessary for cytoskeletal development, is inhibited by Western blotting. Human glioblastoma cell line U87MG cancer cell line by treatment with Sawtooth mabbosa extract and reduction of expression of major proteins involved in cytoskeletal signaling mechanism inhibits cytoskeleton formation, thereby further inhibiting the cell cycle. .

<Experimental Example 6> Inhibitory effect of hexane fraction of teaspoon mushroom on papillary tumor cell metastasis

Human pseudoepidermoid carcinoma cells (U87MG) are known to have a very strong metastatic cancer, which makes it difficult to complete removal by surgical operation and shortens the survival time. In order for cancer cells to show metastasis to other organs, they must have two characteristics: invasiveness and mobility. Invasiveness is a circulatory system called blood flow. It is a circulatory system of blood flow. It is composed of MMP-2 (Matrix matatoproteinase-2), MMP-9 (Matrix matatoproteinase-9) enzyme which dissolves gelatin component existing in blood vessel cell wall and other tissues. Expression and activity are required. And mobility is the ability of cancer cells to move to the point where there is a blood vessel wall so that the cancer cells can easily break away from the tumorigenesis designation. This mobility is a property of the mobility that the surrounding cells move to fill the wound when it is wounded. The western blot analysis shows that the Sawtooth moth extract reduces the expression of MMP-2 and MMP-9 associated with the invasiveness of metastatic U87MG (see FIG. 14). In addition, the transwell invasion assay confirms that U87MG infiltration is actually reduced by reducing the expression of these enzymes (see FIG. 13). Furthermore, it can be confirmed that the wort healing assay inhibits the mobility of U87MG (see FIG. 15).

FIG. 13 is an image obtained by treating the human pseudoepancreatic cell line (U87MG) with the hexane fraction of Production Example 1-2 and confirming invasiveness through a Transwell invasion assay.

14 is an image obtained by Western blotting the amount of expression of MMP-2 and MMP-9 after treating the hexane fraction of Production Example 1-2 with human pseudomyocyte (U87MG).

FIG. 15 is an image obtained by treating the human pseudomonas cell line (U87MG) with the hexane fraction of Production Example 1-2 and confirming the mobility through the Wound healing assay.

As shown in Fig. 13-15, the hexane fraction of the teat mabs was shown to inhibit both the invasiveness and the mobility of human pseudoepithelial cell line (U87MG), and it was confirmed that it has an effect of inhibiting metastasis.

Example of preparation of pharmaceutical agent

The sawtooth extract or sagarchromenol according to the present invention can be formulated into various forms depending on the purpose. Hereinafter, the present invention is not limited thereto, as exemplified by the method of preparing a toothed moth extract or sagarchromenol as an active ingredient according to the present invention.

<Pharmaceutical preparation example 1> Manufacture of powder

Sawtoothmilk extract or Sagar Chromenol 2 g

Lactose 1 g

After mixing the above components, the mixture was packed in an airtight container to prepare a powder.

&Lt; Pharmaceutical Preparation Example 2 > Preparation of tablet

100 mg of Sagamomon extract or Sagar Chromenol

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; Pharmaceutical Preparation Example 3 > Preparation of capsules

100 mg of Sagamomon extract or Sagar Chromenol

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; Pharmaceutical Preparation Example 4 > Preparation of injection

10 mg / ml &lt; / RTI &gt; &lt; RTI ID = 0.0 &gt;

Until dilute hydrochloric acid BP pH 3.5

Sodium chloride BP injected up to 1 ml

The pH of the resulting solution was adjusted to pH 3.5 with diluted hydrochloric acid BP, and the volume was adjusted to the volume using sodium chloride BP for injection Lt; / RTI &gt; The solution was filled in a 5 ml type I ampoule made of transparent glass, sealed in an upper lattice of air by dissolving the glass, sterilized by autoclaving at 120 DEG C for 15 minutes or longer, and an injection solution was prepared.

&Lt; Pharmaceutical Production Example 5 > Production of liquid agent

Sawtooth Extract or Sagar Chromenol 100 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water to dissolve and add lemon fragrance, then the above components were mixed, and purified water was added to adjust the total volume to 100 mL. The solution was filled in a brown bottle and sterilized to prepare a liquid preparation Respectively.

Claims (12)

A pharmaceutical composition for the prophylaxis or treatment of gastric cancer, glioblastoma, or hepatoma containing the hexane fraction obtained from ethanol extract of Sargassum serratifolium as an active ingredient.
delete delete delete delete delete delete The method according to claim 1,
Wherein said cancer is a glioblastoma. delete delete delete delete

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